鉴定南美锥虫病病媒豚鼠血粉的方法。

Juan Carlos Pizarro, David Lucero, Lori Stevens
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引用次数: 0

摘要

背景:在基于 SINE 的 PCR 检测中,使用豚鼠基因组短穿插元件 DNA 的特异性引物组来测试基因组标记在确定 Triatoma infestans 的脊椎动物血餐来源方面的效用:调查包括两项试验。在试验 1 中,在受控条件下,将从玻利维亚丘基萨卡省祖达涅斯(Zudáñez)采集的 36 只昆虫在喂食豚鼠 1-40 小时后冷冻。通过解剖每只昆虫的腹部后部,然后提取 DNA 并进行 PCR 扩增,确认了脊椎动物宿主的种类。试验 2 调查了该技术在野外条件下是否有效。我们分析了从野生豚鼠和人工饲养豚鼠出没的社区的家庭和家庭周边建筑中采集的 34 只昆虫的血食。采集后,昆虫在室温下保存 2 个月,不喂食,然后进行分析:在化验 1 中,36 只被允许吸食豚鼠血液的昆虫均检测出豚鼠 DNA 阳性。豚鼠 DNA 在喂食后 1 小时内和 40 小时内都能被可靠地识别出来。在检测 2 中,34 个样本中有 8 个(23%)用豚鼠特异性引物检测出阳性结果:化验 1 的结果表明,脊椎动物宿主的 DNA 可以在取食血型南美锥虫病病媒 Triatoma infestans 的腹部 1-40 小时后扩增出来。检测 2 的结果证实,该程序适用于从家庭和家庭周边建筑中采集的昆虫,并且可以在取食后至少 2 个月内确定血食的来源。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A method for the identification of guinea pig blood meal in the Chagas disease vector, Triatoma infestans.

A method for the identification of guinea pig blood meal in the Chagas disease vector, Triatoma infestans.

Background: In a SINE-based PCR assay, a primer set specific for guinea pig genome short interspersed elements DNA was used to test the utility of genomic markers for identifying the source of vertebrate blood meals of Triatoma infestans.

Methods: The investigation consisted of two assays. In Assay 1, thirty-six insects, collected from the Province of Zudáñez in Chuquisaca, Bolivia were frozen 1-40 hours after feeding, under controlled conditions, on guinea pigs. The species of the vertebrate host was confirmed from dissection of the posterior part of the abdomen of each insect followed by DNA extraction and PCR amplification. Assay 2 investigated whether the technique worked under field conditions. We analyzed the bloodmeal of 34 insects collected from households and peri-domestic structures from communities where wild and captive guinea pigs occur. After collection, the insects were maintained at room temperature for 2 months without feeding and then analyzed.

Results: In Assay 1, each of the 36 insects allowed to feed on guinea pig blood tested positive for guinea pig DNA. The guinea pig DNA was reliably identified in as little as 1 hour and up to 40 hours after feeding. For Assay 2, 8 out of the 34 samples (23%) showed positive results with guinea pig specific primers.

Conclusion: The results in assay 1 demonstrated that DNA from the vertebrate host can be amplified 1-40 hours post feeding from the abdomen of the blood-feeding Chagas disease vector Triatoma infestans. The results in assay 2 confirmed that the procedure works on insects collected from households and peri-domestic structures and that the source of a blood meal can be determined at least 2 months post feeding.

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